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大规模分离中国仓鼠卵巢细胞的紫外线敏感克隆。

Large-scale isolation of UV-sensitive clones of CHO cells.

作者信息

Busch D B, Cleaver J E, Glaser D A

出版信息

Somatic Cell Genet. 1980 May;6(3):407-18. doi: 10.1007/BF01542792.

Abstract

We have isolated 54 ultraviolet light (UV) sensitive clones of Chinese hamster ovary (CHO) cells, including two from a parent cell line which is hypersensitive to ethyl methanesulfonate (EMS) and is also sensitive to X rays. A replica plating technique was used for the isolation of two of the clones, and a semiautomated technique was used for the isolation of the other 52 clones. We have observed UV sensitization of up to 5-fold in the mutants relative to the parent in terms of the slopes of the survival curves. Seven of the clones were examined for DNA repair competence using a repair replication assay, and all exhibited a DNA repair defect resembling that seen in human mutant xeroderma pigmentosum cells. We have also demonstrated an approximately 9-fold enhancement in the UV mutagenic response of two of the repair replication-defective clones relative to the parent for resistance to ouabain, 6-thioguanine, and 8-azaadenine.

摘要

我们分离出了54个对紫外线(UV)敏感的中国仓鼠卵巢(CHO)细胞克隆,其中两个来自对甲磺酸乙酯(EMS)超敏且对X射线也敏感的亲本细胞系。使用复制平板技术分离了其中两个克隆,使用半自动技术分离了其他52个克隆。就存活曲线的斜率而言,我们观察到突变体相对于亲本的紫外线致敏作用高达5倍。使用修复复制试验检测了其中7个克隆的DNA修复能力,所有克隆均表现出与人类色素性干皮病突变细胞中所见相似的DNA修复缺陷。我们还证明,相对于亲本,两个修复复制缺陷克隆对哇巴因、6-硫鸟嘌呤和8-氮杂腺嘌呤的紫外线诱变反应增强了约9倍。

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